title: Tracking Neospora caninum parasites using chimera monoclonal antibodies against its surface antigen-related sequences (rNcSRS2).
abstract: Neosporosis, an infectious disease of cattle and dogs, causes an abortion in cattle, which has a major damage on the dairy industry worldwide. Tracking of Neospora caninum parasite that is responsible for neosporosis is required for the prevention of this infectious disease. We developed three chimera monoclonal antibodies consist of variable regions of murine antibody and constant regions of human antibody against N. caninum. Recombinant surface antigen-related sequence 2 (rNcSRS2) of N. caninum was expressed in silkworm larvae, and immunized in mice to obtain phage displaying antibody library. Through three rounds of selection, three antibodies, A6, E1 and H3, were isolated and bound to rNcSRS2 with nanomolar to micromolar affinity. In immunofluorescent staining assays, A6 and E1 bound to N. caninum strain Nc-Liv, demonstrating a successful tracking of the parasite. H3 clone bound to rNcSRS2 but not to a truncated protein without glycosylphosphatidylinositol (GPI) anchor domain in the carboxyl terminal. Amino acid sequences of A6 and E1 were similar, but that of H3 differed in the CDR-H1 region, which might be the reason of their difference of affinity. These antibodies are thought to be useful for prevention of cattle from neosporosis.
authors.affiliation: Laboratory of Biotechnology, Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Dong
authors.affiliation: Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Otsuki
authors.affiliation: Laboratory of Biotechnology, Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan; Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Kato
authors.affiliation: Laboratory of Biotechnology, Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan; Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan. Electronic address: acypark@ipc.shizuoka.ac.jp.
authors.forename: Enoch Y
authors.initials: EY
authors.lastname: Park
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000913:Antibodies, Protozoan / Q000276:immunology*; D000953:Antigens, Protozoan / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology*; D012831:Bombyx; D002522:Chlorocebus aethiops; D003048:Coccidiosis / Q000175:diagnosis / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D018688:Neospora / Q000276:immunology / Q000302:isolation & purification*; D019151:Peptide Library; D015800:Protozoan Proteins / Q000276:immunology*; D020349:Surface Plasmon Resonance; D014709:Vero Cells
chemical_list: D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D019151:Peptide Library; D015800:Protozoan Proteins; C115389:SRS2 protein, Neospora caninum
keywords: Antibody; Immunofluorescent staining; Neosporosis; Parasite; Phage display